• Journal of Ginseng Research
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• 제40권4호
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• pp.400-408
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• 2016

Background: Ginsenoside Rh2 (GRh2) is the main bioactive component in American ginseng, a commonly used herb, and its antitumor activity had been studied in previous studies. PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), a serine/threonine protein kinase, is highly expressed in HCT116 colorectal cancer cells. Methods: We examined the effect of GRh2 on HCT116 cells ex vivo. Next, we performed in vitro binding assay and in vitro kinase assay to search for the target of GRh2. Furthermore, we elucidated the underlying molecular mechanisms for the antitumor effect of GRh2 ex vivo and in vivo. Results: The results of our in vitro studies indicated that GRh2 can directly bind with PBK/TOPK and GRh2 also can directly inhibit PBK/TOPK activity. Ex vivo studies showed that GRh2 significantly induced cell death in HCT116 colorectal cancer cells. Further mechanistic study demonstrated that these compounds inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 (ERK1/2) and (H3) in HCT116 colorectal cancer cells. In vivo studies showed GRh2 inhibited the growth of xenograft tumors of HCT116 cells and inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 and histone H3. Conclusion: The results indicate that GRh2 exerts promising antitumor effect that is specific to human HCT116 colorectal cancer cells through inhibiting the activity of PBK/TOPK.

• Journal of Ginseng Research
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• 제39권4호
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• pp.322-330
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• 2015

Background: UV-irradiated keratinocytes secrete various proinflammatory cytokines. UV-induced skin damage is mediated by growth factors and proinflammatory cytokines such as granulocyte macrophage colony stimulating factor (GM-CSF). In a previous study, we found that the saponin of Korean Red Ginseng (SKRG) decreased the expression of GM-CSF in UVB-irradiated SP-1 keratinocytes. In this study, we attempted to find the inhibitory mechanism of SKRG on UVB-induced GM-CSF expression in SP-1 keratinocytes. Methods: We investigated the inhibitory mechanism of SKRG and ginsenosides from Panax ginseng on UVB-induced GM-CSF expression in SP-1 keratinocytes. Results: Treatment with SKRG decreased the expression of GM-CSF mRNA and protein induced by irradiation of UVB in SP-1 keratinocytes. The phosphorylation of ERK was induced by UVB at 10 min, and decreased with SKRG treatment in SP-1 keratinocytes. In addition, treatment with SKRG inhibited the UVB-induced phosphorylation of epidermal growth factor receptor (EGFR), which is known to be an upstream signal of ERK. From these results, we found that the inhibition of GM-CSF expression by SKRG was derived from the decreased phosphorylation of EGFR. To identify the specific compound composing SKRG, we tested fifteen kinds of ginsenosides. Among these compounds, ginsenoside-Rh3 decreased the expression of GM-CSF protein and mRNA in SP-1 keratinocytes. Conclusion: Taken together, we found that treatment with SKRG decreased the phosphorylation of EGFR and ERK in UVB-irradiated SP-1 keratinocytes and subsequently inhibited the expression of GM-CSF. Furthermore, we identified ginsenoside-Rh3 as the active saponin in Korean Red Ginseng.

• Journal of Ginseng Research
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• 제33권4호
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• pp.324-329
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• 2009

G-Rh1에 의한 단핵구 세포주인 U937 세포의 유착조절 능을 조사하여 다음과 같은 결론을 얻었다. 1. G-Rh1은 CD29 항체 (MEM101A) 처리에 의해 유도된 세포-세포간 유착현상을 유의적으로 억제하였다. 2. G-Rh1은 fibronectin처리에 의해 유도된 U937 세포-fibronectin간 유착현상을 유의적으로 억제하였다. 3. G-Rh1은 CD29의 세포표면 발현 수준을 유의적으로 감소시켰다. 최근 활발히 G-Rh1의 약리작용 연구들이 진행되고 있다. 특별히, 본 화합물은 항알러지, 피부염증질환 치료효과, 항암효과 및 여성호르몬 유사기능 등이 있는 것으로 보고된 바있다. CD29-매개성 세포유착과정이 다양한 염증과정, 알러지 반응 및 암세포의 이동 및 전이성과 관련이 있다는 관점에서 볼 때 본 연구결과는 이들 약물이 갖는 치료기전의 하나가 CD29 기능저해에서 비롯될 수 있음을 제시한다고 하겠다. 이후 G-Rh1의 억제 기능에 관한 기전 이해를 위해 세포유착 유도 신호전달 단백질의 활성을 포함한 다양한 분자적 수준에서의 추가적인 실험들을 진행하고 한다.

• 대한약침학회지
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• 제19권3호
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• pp.213-224
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• 2016

Objectives: Ginseng Rh2+ is enzyme-treated ginseng extract containing high amounts of converted ginsenosides, such as compound k, Rh2, Rg3, which have potent anticancer activity. We conducted general and genetic toxicity tests to evaluate the safety of ginseng Rh2+. Methods: An acute oral toxicity test was performed at a high-level dose of 4,000 mg/kg/day in Sprague-Dawley (SD) rats. A 14-day range-finding study was also conducted to set dose levels for the 90-day study. A subchronic 90-day toxicity study was performed at dose levels of 1,000 and 2,000 mg/kg/day to investigate the no-observed-adverse-effect level (NOAEL) of ginseng Rh2+ and target organs. To identify the mutagenic potential of ginseng Rh2+, we conducted a bacterial reverse mutation test (Ames test) using amino-acid-requiring strains of Salmonella typhimurium and Escherichia coli (E. coli), a chromosome aberration test with Chinese hamster lung (CHL) cells, and an in vivo micronucleus test using ICR mice bone marrow as recommended by the Korean Ministry of Food and Drug Safety. Results: According to the results of the acute oral toxicity study, the approximate lethal dose (ALD) of ginseng Rh2+ was estimated to be higher than 4,000 mg/kg. For the 90-day study, no toxicological effect of ginseng Rh2+ was observed in body-weight changes, food consumption, clinical signs, organ weights, histopathology, ophthalmology, and clinical pathology. The NOAEL of ginseng Rh2+ was established to be 2,000 mg/kg/day, and no target organ was found in this test. In addition, no evidence of mutagenicity was found either on the in vitro genotoxicity tests, including the Ames test and the chromosome aberration test, or on the in vivo in mice bone marrow micronucleus test. Conclusion: On the basis of our findings, ginseng Rh2+ is a non-toxic material with no genotoxicity. We expect that ginseng Rh2+ may be used as a novel adjuvant anticancer agent that is safe for long-term administration.

• Journal of Ginseng Research
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• 제44권5호
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• pp.704-716
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• 2020

Background: Ginsenoside Rh2 (GRh2) is a characterized component in red ginseng widely used in Korea and China. GRh2 exhibits a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer properties. However, its effects on Toxoplasma gondii (T. gondii) infection have not been clarified yet. Methods: The effect of GRh2 against T. gondii was assessed under in vitro and in vivo experiments. The BV2 cells were infected with tachyzoites of T. gondii RH strain, and the effects of GRh2 were evaluated by MTT assay, morphological observations, immunofluorescence staining, a trypan blue exclusion assay, reverse transcription PCR, and Western blot analyses. The in vivo experiment was conducted with BALB/c mice inoculated with lethal amounts of tachyzoites with or without GRh2 treatment. Results and conclusion: The GRh2 treatment significantly inhibited the proliferation of T. gondii under in vitro and in vivo studies. Furthermore, GRh2 blocked the activation of microglia and specifically decreased the release of inflammatory mediators in response to T. gondii infection through TLR4/NF-κB signaling pathway. In mice, GRh2 conferred modest protection from a lethal dose of T. gondii. After the treatment, the proliferation of tachyzoites in the peritoneal cavity of infected mice markedly decreased. Moreover, GRh2 also significantly decreased the T. gondii burden in mouse brain tissues. These findings indicate that GRh2 exhibits an antieT. gondii effect and inhibits the microglial activation through TLR4/NF-κB signaling pathway, providing the basic pharmacological basis for the development of new drugs to treat toxoplasmic encephalitis.

• 한국식품과학회지
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• 제39권5호
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• pp.494-499
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• 2007

경북 풍기산 원료삼의 증자 및 건조 조건별 홍삼의 품질 및 기능성을 알아보기 위해 5년 근을 대상으로 증자 조건(2.5, 3.5, 4.5시간)과 건조 조건(열풍건조: $60-65^{\circ}C/24$시간, $40^{\circ}C/3-4$일, 동결 건조: $-80^{\circ}C/56$시간, 원적외선건조: $900W/62^{\circ}C/68$시간) 별로 이화학적 특성과 ginsenoside함량을 비교 분석하였다. 시료의 황색 도와 갈변도는 지근이 본근에 비해 높은 수치를 보였으며, 증자시간 3.5, 4.5, 2.5시간의 순으로 그리고, 원적외선건조, 열풍건조, 동결건조의 순으로 높은 수치를 보였다. 증자 및 건조시료의 가용성 고형분, 총 페놀, 총 플라보노이드, 산성다당체, 전자공여능 등의 특성은 3.5시간 증자와 원적외선 건조가 가장 양호한 결과를 보였다. 조사포닌 함량은 증자시간 및 건조방법에 영향을 받지 않았다. 함량이 높은 $ginsenoside-Rg_1$, -Re, -Rf $-Rb_2$, 등은 증자시간이 길어질수록 감소하였으나 $-Rg_3$ 및 $-Rh_2$의 함량은 유의적으로 증가하여 본근은 3.5시간, 지근은 4.5시간 증자시료에서 가장 높은 함량을 보였다(P<0.01). 증자 시료의 건조방법을 ginsenoside 관점에서 고려한다면 동결건조방법(2,824mg%)이 열풍건조(2,669 mg%)나 원적외선건조(2,667 mg%)에 비해 가장 타당한 것으로 판단되며, $ginsenoside-Rg_3$ 및 $Rh_2$의 함량은 동결건조와 열풍건조 간에 유의적인 차이가 없었다.

• 한국식품과학회지
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• 제38권4호
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• pp.521-525
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• 2006

인삼을 처리온도 및 처리시간을 변수로 하여 열처리한 다음 80% 메탄올로 추출하여 성분 및 생리활성 변화를 분석하였다. 총 폴리페놀 함량은 고온고압처리에 따라 증가하다가 $140^{\circ}C$, 4시간 이후에 감소하였으며, $150^{\circ}C$, 1시간 처리구에서 29.46mg/g으로 가장 높은 함량을 나타내었다. 총 플라보노이드 함량은 고온고압처리에 따라 증가하다가 $150^{\circ}C$에서는 2시간 처리구에서 4.75mg/g으로 가장 높은 함량을 나타내었다. $IC_{50}$은 처리온도와 시간이 증가할수록 감소하여 항산화활성이 무처리구(17.68mg/g)보다 증가한 것을 알 수 있었으며, 가장 활성이 높은 처리구는 $140^{\circ}C$, 3시간 처리구로 0.22mg/g으로 나타났다. 4년근 인삼의 조사포닌 함량은 1.18%이었으며, 고온고압처리에 따라 ginsenoside는 대부분 처리온도가 높아질수록, 처리시간이 길어질수록 감소하는 경향을 보였다. $Rg_1$, Re, $Rb_2,\;Rb_3$은 비교적 낮은 온도에서는 안정하였으나 $130^{\circ}C$ 이상의 온도에서는 불안정하여 감소하였다. Rf는 열처리에 비교적 안정하였으며, $Rg_3,\;Rh_2$는 고온고압처리에 의해 새로이 생성되거나 함량이 증가하여 최대 생성 조건은 $130^{\circ}C$에서는 4-5시간, $140^{\circ}C$에서는 2-4시간, $150^{\circ}C$에서는 2시간으로 나타났다.

• Journal of Ginseng Research
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• 제19권3호
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• pp.254-259
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• 1995

The content and composition of saponin compounds of Panax species were analyzed according to their species, region and processing type of red and white ginseng. The species employed were Korean-, Chinese-, Japanese red ginsengs, and Korean white ginseng of Panax ginseng, American- and Canadian ginsengs of Panax quinquefolium, and Panax notoinseng. Twelve main saponin components in the ginseng were identified and quantified using TLC and HPLC. All three species had remarkably different content and composition. However, within each species they were similar. Twelve major ginsenosides were determined in P. ginseng, eight in p. quinquefolium, and six in P. notoginseng. Of the components of P ginseng Rf, $Rh_1$, $Rh_2$ and Ra were not detected in P quinquefolium, and $Rb_2$, Rc, Rf, $Rh_2$, Ra and Ro not detected in P. notoinseam. Crude saponin content and protopanaxadiol/protopanaxatriol saponin ratio were compared. They were 4.81~5.24% and 1.27~ 1.45 in p. ginsengs, 7.01~7.25% and 2.12~ 2.15 in p. quinquefolium, 9.80% and 0.99 in P. notoineng. The prosapogenin and sapogenin content were different among the Panax species.

• Journal of Ginseng Research
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• 제45권2호
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• pp.295-304
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• 2021

Background: Acute promyelocytic leukemia (APL) is a hematopoietic malignancy driven by promyelocytic leukemia-retinoic acid receptor A (PML-RARA) fusion gene. The therapeutic drugs currently used to treat APL have adverse effects. 20(S)-ginsenoside Rh2 (GRh2) is an anticancer medicine with high effectiveness and low toxicity. However, the underlying anticancer mechanisms of GRh2-induced PML-RARA degradation and apoptosis in human APL cell line (NB4 cells) remain unclear. Methods: Apoptosis-related indicators and PML-RARA expression were determined to investigate the effect of GRh2 on NB4 cells. Z-VAD-FMK, LY294002, and C 87, as inhibitors of caspase, and the phosphatidylinositol 3-kinase (PI3K) and tumor necrosis factor-α (TNF-α) pathways were used to clarify the relationship between GRh2-induced apoptosis and PML-RARA degradation. Results: GRh2 dose- and time-dependently decreased NB4 cell viability. GRh2-induced apoptosis, cell cycle arrest, and caspase3, caspase8, and caspase9 activation in NB4 cells after a 12-hour treatment. GRh2-induced apoptosis in NB4 cells was accompanied by massive production of reactive oxygen species, mitochondrial damage and upregulated Bax/Bcl-2 expression. GRh2 also induced PML/PML-RARA degradation, PML nuclear bodies formation, and activation of the downstream p53 pathway in NB4 cells. Z-VAD-FMK inhibited caspase activation and significantly reversed GRh2-induced apoptosis and PML-RARA degradation. GRh2 also upregulated TNF-α expression and inhibited Akt phosphorylation. LY294002, an inhibitor of the PI3K pathway, enhanced the antitumor effects of GRh2, and C 87, an inhibitor of the TNF-α pathway, reversed NB4 cell viability, and GRh2-mediated apoptosis in a caspase-8-dependent manner. Conclusion: GRh2 induced caspase-dependent PML-RARA degradation and apoptosis in NB4 cells via the Akt/Bax/caspase9 and TNF-α/caspase8 pathways.

• 생약학회지
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• 제47권1호
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• pp.73-78
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• 2016

The purpose of this study is to develop a new preparation process of ginseng (Panax ginseng) flower buds extracts featuring high concentration of ginsenosides Rg2, Rg3, Rg5, F4 and Rh1, red ginseng special components. Chemical transformation from ginseng saponin glycosides to prosapogenin was analyzed by the HPLC. Extracts of ginseng flower buds were processed under several treatment conditions of ultrasonication (at $100^{\circ}C$). The results showed that the quantity of ginsenoside Rg6 increased by over 8.8% at the 16 hours of ultrasonication. Ginseng flower buds ethanol extract compared with other process times. The result of UGF-16 indicates that the ultrasonication processed ginseng flower buds extracts (at $100^{\circ}C$) treated for 16 hours produced the highest amount of ginsenoside F4 (8.833%), Rg3 (2.230%), Rg5 (2.339%) and Rg2 (1.002%).